Ridlen J S, Skotty D R, Kissinger P T, Nieman T A
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana 61801, USA.
J Chromatogr B Biomed Sci Appl. 1997 Jul 4;694(2):393-400. doi: 10.1016/s0378-4347(97)00138-2.
Erythromycin is determined in both urine and plasma samples using microbore reversed-phase liquid chromatography with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)] electrogenerated chemiluminescence (ECL) detection. Ru(bpy)3(2+) is included in the mobile phase thus eliminating band broadening caused by post-column reagent addition. Extra column band broadening is an important concern in microbore liquid chromatography due to the small peak volumes. Erythromycin was studied in both water and biological samples. The detection limit for erythromycin in standards is 0.01 microM or 50 fmol injected with a S/N of 3 and a linear working range that extends four orders of magnitude. Human urine and blood plasma were also studied. Urine samples were diluted and filtered before injection. Ultrafiltration was used to remove protein from blood plasma samples prior to injection. Erythromycin was selectively detected in the body fluid samples without any further sample preparation. The detection limits obtained for erythromycin in urine and plasma are 0.05 and 0.1 microM, respectively, for 5 microl injected on a 150x1 mm I.D. C18 column.
采用带有三（2,2'-联吡啶）钌（II）[Ru(bpy)₃(2+)] 电化学发光（ECL）检测的微径反相液相色谱法测定尿液和血浆样品中的红霉素。流动相中包含Ru(bpy)₃(2+)，从而消除了柱后添加试剂导致的峰展宽。由于峰体积小，柱外峰展宽是微径液相色谱中的一个重要问题。对红霉素在水和生物样品中的情况进行了研究。标准品中红霉素的检测限为0.01 μM或进样50 fmol，信噪比为3，线性工作范围扩展四个数量级。还对人尿液和血浆进行了研究。尿液样品在进样前进行稀释和过滤。血浆样品在进样前采用超滤法去除蛋白质。无需进一步样品制备即可在体液样品中选择性检测到红霉素。在150×1 mm内径的C18柱上进样5 μl时，尿液和血浆中红霉素的检测限分别为0.05和0.1 μM。
